Biomedical subjects

N Terada

Publications and source records attributed to N Terada.

At least 163 records · Page 9Linked to original sources

PubMed1997

The effects of eotaxin on the surface adhesion molecules of endothelial cells and on eosinophil adhesion to microvascular endothelial cells.

Eosinophil recruitment occurs in tissues as the result of allergic diseases. Human eotaxin is thought to be specific to eosinophils. In this study, we examined the effects of human eotaxin on the expression of adhesion molecules on nasal microvascular endothelial cells and on eosinophil adhesion to endothelial cells. Eotaxin upregulated the expression of ICAM-1 and VCAM-1 on human nasal mucosal microvascular endothelial cells (HMMEC), but not human umbilical vein endothelial cells (HUVEC). The eotaxin-induced eosinophil adhesion to HMMEC was increased at 10 ng/ml and significantly increased at the concentration of 100 ng/ml. On HUVEC, however, eotaxin did not induce increases of eosinophil adhesion. Anti-ICAM-1 and anti-VCAM-1 mAbs significantly decreased eotaxin-induced eosinophil adhesion. These results suggest that eotaxin regulates eosinophil accumulation to the nasal mucosa through its effect on the adhesion molecules on microvascular endothelial cells.

Adult↗

PubMed1997

Overexpression of wild type p70 S6 kinase interferes with cytokinesis.

p70 S6 kinase (p70s6k) is a serine/threonine kinase which is activated through an unidentified pathway by mitogenic stimuli. Here we demonstrate that stable- and transient-overexpression of wild type (WT)-p70s6k results in perturbation of cytokinesis in NIH3T3 cells. Cells overexpressing WT-p70s6k demonstrated a slow growth rate and lower viability, whereas cells overexpressing a mutant T229A-p70s6k had a similar growth rate and viability as those of parental NIH3T3 cells. Moreover, WT-p70s6k cells demonstrated several abnormalities in cellular morphology; both the cytoplasm and the nuclei were large and a proportion of the cells was multinucleated. Flow cytometric analysis of propidium iodide-stained cells demonstrated that about 50% of the cells had 4N or greater content of DNA. When cell division in WT-p70s6k cells was monitored by time-lapse video microscopy, cytokines was often incomplete after mitosis, producing binucleated cells. Addition of rapamycin to WT-p70s6k cells, which inactivates endogenous and transfected p70s6k, failed to reverse any of the morphological abnormalities. These indicate that overexpression of p70s6k but not increases in activity of p70s6k, is responsible for the phenotype. A molecule which interacts with p70s6k may be involed in this regulation of cytokinesis.

3T3 Cells↗

PubMed1997

Mast cell tumor necrosis factor alpha production is regulated by MEK kinases.

Mast cells synthesize and secrete specific cytokines and chemokines which play an important role in allergic inflammation. Aggregation of the high-affinity Fc receptor (FcepsilonRI) for immunoglobulin E (IgE) in MC/9 mouse mast cells stimulates the synthesis and secretion of tumor necrosis factor alpha (TNF-alpha). FcepsilonRI aggregation activates several sequential protein kinase pathways, leading to increased activity of extracellular signal-regulated kinases (ERKs), c-Jun amino-terminal kinases (JNKs), and the p38 mitogen-activated protein (MAP) kinase. Inhibition of ERKs with the compound PD 098059 had little effect on FcepsilonRI-stimulated TNF-alpha production. Aggregation of FcepsilonRI stimulated MEK kinase 1 (MEKK1) activity, which activates JNK kinase (JNKK), the kinase that phosphorylates and activates JNKs. Expression of activated MEKK1 (DeltaMEKK1) in MC/9 cells strongly stimulated JNK activity but only weakly stimulated p38 activity, and it induced a large activation of TNF-alpha promoter-regulated luciferase gene expression. Inhibitory mutant JNK2 expressed in MC/9 cells significantly blunted FcepsilonRI stimulation of TNF-alpha promoter-driven luciferase expression. Wortmannin, an inhibitor of phosphatidylinositol 3-kinase, diminished FcepsilonRI-mediated TNF-alpha synthesis, significantly blunted JNK activation and TNF-alpha promoter-driven luciferase expression, and only weakly inhibited p38 kinase activation. Inhibition of NFkappaB activation resulting from DeltaMEKK1 expression or FcepsilonRI stimulation did not affect TNF-alpha promoter-driven luciferase expression. Our findings define a MEKK-regulated JNK pathway activated by FcepsilonRI that regulates TNF-alpha production in mast cells.

Androstadienes↗

PubMed1997

Eosinophil adhesion regulates RANTES production in nasal epithelial cells.

Among the many known chemotactic factors for eosinophils, the proinflammatory chemokine RANTES is particularly important, because it is potently and selectively chemotactic for eosinophils. Throughout the process of the migration of eosinophils from the blood vessels into the nasal cavity, eosinophil functions are assumed to be regulated by surface adhesion molecules. Conversely, the messages conferred by the eosinophils to the endothelial and epithelial cells are also of great interest. In the present study, we showed that eosinophil adhesion to human nasal epithelial cells (HNECs) inhibits RANTES production in HNECs. Eosinophils were isolated from peripheral blood obtained from patients with allergic rhinitis. Human mucosal microvascular endothelial cells and HNECs were isolated from human nasal mucosa specimens. After stimulation of the HNECs in the presence of eosinophils, the secretion of RANTES, induced by a combination of TNF-alpha and IFN-gamma, appeared to have decreased. The amount of the decrease was a function of the number of involved eosinophils. On the other hand, the presence of eosinophils did not affect RANTES production by the endothelial cells. After pretreatment of the eosinophils with anti-CD18 mAb or coculture with HNECs in Transwell culture inserts, these cells did not inhibit the TNF-alpha- and IFN-gamma-induced RANTES production. These results were virtually identical with those observed on RANTES mRNA expression. The adhesion of eosinophils to HNECs plays a key role in the inhibition of RANTES production. Our data indicate that a certain established system causes the signal transfer from eosinophils to HNECs to inhibit RANTES production, thus decreasing the eosinophil infiltration.

Adult↗

PubMed1997

Rapamycin potentiates dexamethasone-induced apoptosis and inhibits JNK activity in lymphoblastoid cells.

The immunosuppressant rapamycin (RAP) potentiated apoptosis of the murine T lymphoblastoid cell line S49 induced by dexamethasone (DEX), while RAP by itself did not induce apoptosis of the cells. FK506, in contrast, had no effect on DEX-induced apoptosis; moreover, an excess of FK506 reversed the potentiation of apoptosis by RAP, indicating that RAP exerts its effects through binding to FKBP. Both RAP and FK506 enhanced the MMTV promoter activity by dexamethasone, suggesting that the potentiation of apoptosis is not likely explained by the selective enhancement of transcriptional activity of the glucocorticoid receptor. Of interest, the basal activity of c-Jun kinase (JNK), whose activation has been recently suggested to be involved in cell survival signals in lymphocytes, was reduced by RAP in S49 cells. The reduction of JNK activity by RAP was reversed by the addition of an excess of FK506. In summary, we demonstrate for the first time that RAP has the ability to inhibit JNK activity in lymphocytes where the drug enhances apoptosis.

3T3 Cells↗

PubMed1997

Vesnarinone inhibits adenosine uptake in endothelial cells, smooth muscle cells and myocytes, and mediates cytoprotection.

Vesnarinone is a novel synthetic inotropic agent. Recently, it has been reported that vesnarinone inhibits adenosine uptake in the B-lymphocytoid cell line. Since extracellular adenosine is cardioprotective, we examined whether vesnarinone inhibits adenosine uptake in cells constituting the cardiovascular system. 1 microCi of -3H-adenosine was added to cells of the myocyte cell line (C2C12), human coronary smooth muscle cell line (HCASMC), human and bovine coronary endothelial cell lines (HCAEC and BCAEC), bovine arterial endothelial cell line (BAEC), and human umbilical venous endothelial cell line (HUVEC). After 10 s-5 min, cells were separated from free [3H]adenosine, and the radioactivity was measured. When 0.1-100 microM of vesnarinone was added to each cell line, the uptake of adenosine was inhibited dose-dependently {% inhibition of -3H-adenosine uptake at 10 and 30 microM of vesnarinone: 14 and 33% (C2C12), 47 and 72% (HCASMC), 37 and 58% (HCAEC), 42 and 68% (BCAEC), 19 and 68% (BAEC), 29 and 59% (HUVEC)}. The cellular viability of HCAEC exposed to 60 min of hypoxia and 60 min of reoxygenation increased from 34+/-5 to 67+/-6% (Trypan blue exclusion test) and 23+/-5 to 78+/-6% (LDH release), which was completely blunted by 8-sulfophenyltheophylline, an adenosine receptor antagonist, and was partially blunted by alpha,beta-methyleneadenosine 5'-diphosphate, an inhibitor of ecto-5'-nucleotidase. We also found that vesnarinone is cytoprotective against hypoxia and reoxygenation in C2C12 and HCASMC. We conclude that vesnarinone inhibits the uptake of adenosine in cardiovascular cells, which contributes to cytoprotection.

Adenosine↗

PubMed1997

Dystrophin in rod spherules; submembranous dense regions facing bipolar cell processes.

It is known that the retina contains the protein dystrophin in the ribbon synapse, but the ultrastructural analysis is not yet fully elucidated. Our previous study reported that dystrophin is localized under the rod cell membranes in rat retinas. In the present study, we have investigated the relationship between dystrophin-rich regions of rod cell membranes and other neuronal processes in mouse retinas with a monoclonal antibody raised against the human dystrophin C-terminus. Immunoblotting, immunofluorescence stainings, and immunoelectron microscopy were employed. Immunoblotting analysis indicated that mouse retinas possessed some of the dystrophin isoforms of approximately 260 kDa, 140 kDa, and 70 kDa molecular weight. Confocal images showed a punctate appearance in the outer plexiform layer, as previously described. Immunoelectron microscopy showed that dystrophin immunoreactive products were always observed at submembranous dense regions of the rod spherule abutting bipolar processes. These results suggest that retinal dystrophin may be closely involved in signal transmission from rods to bipolar cells.

Animals↗

PubMed1997

A new type of "bioartificial" nerve graft for bridging extended defects in nerves.

In the rat sciatic nerve, a gap of around 10 mm in nerve continuity seems to be the maximal distance which can be successfully repaired by silicone tubes. In this study we tested if a new artificial nerve graft, composed of eight polyamide filaments (diameter 250 microns) placed inside silicone tubes (1.8 mm inner diameter), could be used to bridge an extended gap (15 mm) in rat sciatic nerve. Silicone tubes containing eight polyamide sutures were found to support regeneration across such a gap. After 4 weeks sensory fibres had bridged the gap and grown into the distal nerve segment as revealed by a positive pinch reflex test as well as positive staining for neurofilaments in the distal nerve segment. Myelinated axons could be observed in the tissue matrix formed in between and peripheral to the synthetic filaments along the whole length of the tube. In contrast, when silicone tubes without filaments were used to bridge the 15 mm gap, the tubes contained only fluid or in two cases a thin tissue strand. No positive pinch reflex response was elicited in the nerve segment distal to such a tube. We conclude that the new artificial nerve graft can be used to support regeneration across extended gaps in nerves.

Animals↗

PubMed1997

The role of macrophages in bioartificial nerve grafts based on resorbable guiding filament structures.

A 10 mm gap in a rat sciatic nerve was bridged by a bioartificial nerve graft consisting of a silicone tube containing seven longitudinally placed filaments made of non-resorbable material (polyamide [Ethilon]) or resorbable materials (polydioxanon [PDS], polyglactin [Vicryl] or catgut). The purpose was to study the tissue reaction induced by the four different types of materials. At 4 weeks an immunocytochemical technique, using ED1 and ED2 monoclonal antibodies, was used to study the presence and location of macrophages. A large number of macrophages were found accumulating on the surface of catgut and polyglactin, while few were found on the surface of polyamide and polydioxanon filaments. It is concluded that the cell layers on the filament surface mainly consisted of ED1 positive cells and their thickness depends on the filament materials.

Journal Article↗

PubMed1997

An immunocytochemical study of changes in the human erythrocyte membrane skeleton produced by stretching examined by the quick-freezing and deep-etching method.

A quick-freezing and deep-etching method in combination with erythrocyte splitting was used to examine the cytoplasmic aspect of whole-mount human erythrocyte membranes. Various external forces induced alterations in membrane skeletal organisation during the splitting procedure. The initial change was elongation in the peripheral part of the membrane skeleton, examined by immunostaining with a monoclonal antispectrin antibody. Under severe stretching conditions, a linear rearrangement of filamentous components was evident; these were disposed parallel to the rim of the erythrocyte, while the central part of the concavity exhibited a more compacted structure. These changes resulted in a different distribution of membrane skeletal components between central rigid and peripheral flexible areas in biconcave erythrocytes. It is suggested that the reversible membrane skeletal changes in the flexible areas which resist the external forces are important for maintaining the normal framework of biconcave human erythrocytes.

Antibodies, Monoclonal↗

PubMed1997

Electron microscopic study of erythroblastic islands obtained by 'tissue-stamp culture' method.

A new 'tissue-stamp culture' method was developed for stamping proliferating erythroblasts of mouse spleens on collagen-coated coverslips after inducing haemolytic anaemia by administration of 1-acetyl-2-phenylhydrazine, and then adherent splenic cells were cultured for a few days. We could obtain many erythroblastic islands, where cultured erythroblasts were located over macrophages and were proliferated synchronously for 10-30 h, and then the erythroblasts were differentiated and enucleated after 30-50 h in the presence of erythropoietin. To observe three-dimensional structures of the erythroblastic islands, a scanning electron microscope was used for the cultured cells treated with critical point-drying method. Immature wrinkled erythroblasts with many micropinocytic pits were attached to the central area of the flattened macrophages with many cytoplasmic projections, though matured erythroblasts were localized on their peripheral areas. Moreover, cytoplasmic projections of underlying macrophages, which were attached to the matured erythroblasts, were decreased in number. At a late stage, deep cytoplasmic invaginations of erythroblasts observed at a middle stage became shallow after their enucleation and flattened to form their concave shapes. This 'tissue-stamp culture' system would be useful for studying specific interaction between stromal macrophages and haematopoietic cells.

Anemia, Hemolytic↗

PubMed1997

Diesel exhaust particulates enhance eosinophil adhesion to nasal epithelial cells and cause degranulation.

Diesel exhaust particulates (DEP) are a common air pollutant from diesel-engine-powered car exhaust and are thought to cause chronic airway diseases. On the other hand, eosinophils are major components of allergic inflammatory disorders such as asthma, nasal allergy and atopic dermatitis. We examined the effects of DEP and DEP extract (extract of polyaromatic hydrocarbons) on eosinophil adhesion, survival rate and degranulation. Eosinophils, human mucosal microvascular endothelial cells (HMMECs) and human nasal epithelial cells (HNECs) were preincubated in the presence or absence of DEP and DEP extract. 35S-labeled eosinophils were allowed to adhere to monolayers of HMMECs and HNECs. After washing, 35S radioactivity was determined and numbers of adherent eosinophils were calculated using each standard curve. The effects of DEP and DEP extract on eosinophil survival rate and degranulation were also determined. Although neither DEP nor DEP extract affected the adhesiveness of HMMECs and HNECs to eosinophils, 5 ng/ml of DEP extract and 50 ng/ml of DEP extract each significancy increased eosinophil adhesiveness to HNECs (134+/-9 and 143+/-8%, respectively; p<0.01 vs. control), but neither effected eosinophil adhesiveness to HMMECs. DEP extract also induced eosinophil degranulation without changing the eosinophil survival rate. Given that eosinophil-derived lipid mediators and toxic proteins play important roles in the development of nasal allergy, the above findings strongly suggest that DEP plays an important role in promoting the nasal hypersensitivity induced by enhanced eosinophil infiltration of epithelium and eosinophil degranulation.

Adult↗

PubMed1997

Ultrastructural study of axonal cytoskeletons in the optic nerve damaged by acutely elevated intraocular pressure using the quick-freezing and deep-etching technique.

The aim of this study was to examine the ultrastructure of axonal cytoskeletons in guinea pig optic nerve damage induced by acutely elevated intraocular pressure (IOP) employing the quick-freezing and deep-etching method. An IOP of 60 mm Hg was maintained for 4 h. The unmyelinated axoplasm of the optic nerve at the normal IOP was filled with longitudinally oriented neurofilaments and bundled microtubules, which were cross-linked by numerous cross-bridges. Membranous organelles, such as mitochondria and vesicles, were scattered in the axoplasm. These organelles were similarly interconnected with microtubules or neurofilaments by the cross-bridges. The unmyelinated axoplasm of the optic nerve following acutely elevated IOP was shown to include intraaxonal collections of various membranous organelles and reduction of microtubules and neurofilaments. The cross-bridges linking microtubules, neurofilaments and membranous organelles in such axoplasm appeared to be dramatically reduced in number. Thus, it is suggested that the ultrastructural changes of axonal cytoskeletons in some optic nerves following elevated IOP might include the reduction of microtubules, neurofilaments and their associated cross-bridges.

Animals↗

PubMed1997

Bioartificial nerve grafts based on absorbable guiding filament structures--early observations.

Gaps 10 mm wide in the sciatic nerves of 64 rats were bridged by bioartificial nerve grafts consisting of a silicone tube containing seven longitudinally placed filaments made of non-absorbable, (polyamide [Ethilon]) or absorbable, material (polydioxanone [PDS], polyglactin [Vicryl], and catgut). The purpose was to study the organisation of axonal growth inside the tube along such filaments. After two and four weeks histological techniques were used to study the contents of the tube and at four weeks immunohistological techniques were used to confirm the presence of axons distal to the tube. In all experimental groups axons had traversed the tube and reached the distal segment after four weeks. Inside the tube axons were organised in multiple minifascicles in all groups, but there were no axons growing in direct contact with the filaments. We conclude that resorbable filaments placed inside a silicone tube do not disturb axonal growth across the tube.

Absorption↗

PubMed1997

Membrane skeletons in avian erythrocytes as revealed by the quick-freezing and deep-etching method.

Ultrastructure of chicken erythrocytes were examined by the quick-freezing and deep-etching (QF-DE) method. Some erythrocytes were fixed with paraformaldehyde and prepared with erythrocyte-splitting method or saponin treatment to remove soluble proteins before quick-freezing. Others were prepared in the cytosol buffer with the erythrocyte-splitting method to obtain natural state of cytoskeletons. Non-expanding membrane skeletons were highly condensed on the cytoplasmic side of lipid membrane in the paraformaldehyde-fixed specimens. Under unilateral extension of the specimens, long stretched filaments were connected alternately with condensed filamentous or granular structures under erythrocyte membranes. As the membrane skeletons got closer to the marginal bands, they become more dense network structures. Moreover, in the fresh unfixed specimens, dense networks of filaments were localized underlying erythrocyte membranes in a relatively intact state. Fine filaments connected the marginal microtubule bands to the cytoplasmic sides of erythrocyte membranes. The different distribution of each cytoskeletal component and the association of these structures may support the elliptocytic shape of chicken erythrocytes and resist the dynamic circumstance.

Animals↗

PubMed1997

Effects of methylcobalamin on the proliferation of androgen-sensitive or estrogen-sensitive malignant cells in culture and in vivo.

Methylcobalamin is one of the coenzymatically active cobalamin derivates and required for the activity of the cytoplasmic enzyme methionine synthetase catalyzing the methylation of homocysteine into methionine. The effect of methylcobalamin on the proliferation of malignant cells has been examined. Methylcobalamin inhibited the proliferation of androgen-sensitive SC-3 cells (a cloned cell line from Shionogi mouse mammary tumor, SC115) in culture at the concentration of 100-300 micrograms/ml. An inhibitory activity of methylcobalamin on the proliferation was also observed in other cell lines (estrogen-sensitive B-1F cells from mouse Leydig cell tumor and MCF-7 cells from human mammary tumor) at the concentration of 500 micrograms/ml. Moreover, large doses of methylcobalamin injected intraperitoneally (100 mg/kg body weight/day) were non-toxic and suppressed the tumor growth of SC115 and B-1F cells in mice fed a vitamin B12 deficient diet. These results show that methylcobalamin inhibits the proliferation of malignant cells in culture and in vivo and propose the possibility of methylcobalamin as a candidate of potentially useful agents for the treatment for some malignant tumors.

Androgens↗

PubMed1997

Immunocytochemical study of dystrophin localization in cone cells of mouse retinas.

PURPOSE: Previously, the authors reported that dystrophin was observed under the rod cell membranes in rat retinas. However, it was not determined whether dystrophin is located in cone cells. In the current study, the authors clarify dystrophin localization in cone cells of mouse retinas. METHODS: Immunoblotting, confocal laser scanning microscopy, and immunoelectron microscopy were used to investigate retinal dystrophin with a monoclonal antibody raised against the human dystrophin C-terminus. RESULTS: Immunoblotting analysis showed some immunoreactive bands from retinal extracts. Confocal images indicated two different immunostaining patterns: One was a tiny dot, and the other was a larger, aggregated dot. Immunoelectron microscopy revealed that retinal dystrophin was localized in cone cells as well as in rod cells. CONCLUSIONS: Retinal dystrophin is a common component of cone and rod cells and probably is related to the physiological function of photoreceptor cells.

Animals↗

PubMed1997

[Effects of TNM extension and histopathological type of the tumor on long-term survival rates of parotid and submandibular gland cancer].

The effects of TNM extension and histopathological type of the tumor on 5 and 10-year survival rates were analysed in 1,147 cases of parotid cancer and 271 cases of submandibular gland cancer which had been registered from 152 institutes throughout Japan since 1978. Advance of stage decreased the prognosis significantly, and the decrease was more pronounced in submandibular gland cancer. Facial paralysis also decreased the prognosis of parotid cancer significantly. The prognosis was also affected significantly by the histopathological type of the tumor. The 5-year survival rates of adenocarcinoma, epidermoid carcinoma and undifferentiated carcinoma were only about 30% in parotid cancer and about 20% in submandibular gland cancer. In treatment of salivary gland cancer, the therapeutic approach should be individualized in accordance with the tumor extension and its histopathological type. Improvement of accuracy in preoperative diagnosis of salivary gland cancer is of the utmost importance.

Adenocarcinoma↗